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The etiological agent for infective endocarditis (IE), a life-threatening disease, is usually gram-positive bacteria. However, gram-negative bacteria can rarely cause IE and 4% of cases are associated with morbidity and mortality. This study aimed to characterize Escherichia coli and Klebsiella pneumoniae isolates from the blood of patients with IE. The characteristics of blood isolates were compared with those of urinary isolates from patients with urinary tract infections (UTIs). The results of this study revealed that K. pneumoniae isolates from patients with IE were phylogenetically related to those from patients with UTI. Additionally, the resistance phenotype, resistance gene, virulence gene, and plasmid profiles were similar between the blood and urinary isolates. The isolates belonging to the sequence types (STs) 76, 36, 101 (K. pneumoniae), and 69 (E. coli) are reported to be associated with drug resistance. The Enterobacteriaceae isolates from patients with IE did not produce extended-spectrum β-lactamase or carbapenemase. Additionally, this study investigated the virulence phenotype, biofilm formation ability, and the ability to adhere to the epithelial cells in vitro of the isolates. The isolates from patients with IE exhibited weaker biofilm formation ability than the urinary isolates. All isolates from patients with IE could adhere to the renal epithelial cells. However, three isolates from patients with UTIs could not adhere to the epithelial cells. The closely related K. pneumoniae isolates (648, KP1, KP2, KP3, and KP4) could not form biofilms or adhere to the epithelial cells. In summary, the molecular analysis revealed that the genetic characteristics of IE-causing K. pneumoniae and E. coli were similar to those of UTI-causing isolates. These isolates belonged to the STs that are considered treatable. Genetically similar isolates did not exhibit the same virulence phenotype. Thus, these non-hypervirulent clones must be monitored as they can cause complex infections in susceptible hosts., (© 2021. The Author(s).)

Background: The competence of enteroaggregative Escherichia coli (EAEC) to adhere to the intestinal epithelium of the host is a key role to the colonization and disease development. The virulence genes are crucial for EAEC pathogenicity during adherence, internalization and persistence in the host. The overwhelming majority of antigen encounters in a host occurs on the intestine surface, which is considered a part of innate mucosal immunity. Intestinal epithelial cells (IECs) can be activated by microorganisms and induce an immune response., Objective: The present study investigated the interaction of invasive EAEC strains with T84 intestinal epithelial cell line in respect to bacterial invasiveness, persistence and cytokines production., Methods: We evaluated intracellular persistence of invasive EAEC strains (H92/3, I49/3 and the prototype 042) and production of cytokines by sandwich ELISA in T84 cells upon 24 hours of infection., Results: The survival rates of the prototype 042 was 0.5x103 CFU/mL while survival of I49/3 and H92/3 reached 3.2x103 CFU/mL and 1.4x103 CFU/mL, respectively. Infection with all EAEC strains tested induced significant amounts of IL-8, IL-6 and TNF-α compared to uninfected T84 cells., Conclusion: These data showed that infection by invasive EAEC induce a proinflammatory immune response in intestinal epithelial T84 cells.

Background: The diarrheal syndrome is considered a serious public health problem all over the world and is considered a major cause of morbidity and mortality in developing countries. The high incidence of enteroaggregative Escherichia coli in diarrheal syndromes classified as an emerging pathogen of gastrointestinal infections. After decades of study, your pathogenesis remains uncertain and has been investigated mainly using in vitro models of adhesion in cellular lines., Objective: The present study investigated the interaction of enteroaggregative Escherichia coli strains isolated from childhood diarrhea with rabbit ileal and colonic mucosa ex vivo, using the in vitro organ culture model., Methods: The in vitro adhesion assays using cultured tissue were performed with the strains co-incubated with intestinal fragments of ileum and colon over a period of 6 hours. Each strain was tested with three intestinal fragments for each region. The fragments were analysed by scanning electron microscopy., Results: Through scanning electron microscopy we observed that all strains adhered to rabbit ileal and colonic mucosa, with the typical aggregative adherence pattern of "stacked bricks" on the epithelium. However, the highest degree of adherence was observed on colonic mucosa. Threadlike structures were found in greater numbers in the ileum compared to the colon., Conclusion: These data showed that enteroaggregative Escherichia coli may have a high tropism for the human colon, which was ratified by the higher degree of adherence on the rabbit colonic mucosa. Finally, data indicated that in vitro organ culture of intestinal mucosa from rabbit may be used to elucidate the enteroaggregative Escherichia coli pathogenesis.